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Exposure to chronic hyperglycemic conditions results in Ras-related C3 botulinum toxin substrate 1 (Rac1)-mediated activation of p53 and ATM kinase in pancreatic β-cells

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Abstract

Chronic hyperglycemia (HG) promotes pancreatic islet dysfunction which leads to the onset of T2DM. This study is aimed at defining regulatory roles of Rac1, a small G-protein, in the activation of p53 and ATM kinase in pancreatic β-cells, under the duress of HG conditions. We report significant stimulatory effects of HG (20 mM; 24 h) on p53 activation in INS-1 832/13 cells, normal rodent and human islets. Pharmacological inhibition of Rac1 (EHT1864 or NSC23766) significantly suppressed HG-induced p53 activation in INS-1 832/13 cells and rat islets, suggesting novel roles for this small G-protein in the activation of p53. Inhibition of Rac1 geranylgeranylation with simvastatin or GGTI-2147, significantly attenuated HG-induced p53 activation, suggesting requisite roles for this signaling step in HG-mediated effects on β-cells. HG-induced p53 activation was also suppressed by SB203580, a known inhibitor of p38MAPK. Additionally, we observed increased activation of ATM kinase under HG conditions, which was blocked in presence of EHT1864. Furthermore, pharmacological inhibition of ATM kinase (KU55933) reduced activation of ATM kinase, but not p53, suggesting that HG-mediated activation of p53 and ATM could represent independent pro-apoptotic events. In conclusion, these data indicate that sustained activation of Rac1-p38MAPK signaling axis leads to activation of p53 leading to β-cell dysfunction under the duress of chronic hyperglycemic conditions.

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Abbreviations

ATM kinase:

Ataxia telangiectasia mutated kinase

GAPs:

GTPase-activating proteins

GEFs:

Guanine nucleotide exchange factors

GTP:

Guanosine triphosphate

GGTI:

Geranylgeranyl transferase inhibitor

GSIS:

Glucose-stimulated insulin secretion

HG:

High glucose

LG:

Low glucose

Nox2:

NADPH oxidase 2

p38MAPK:

p38 mitogen activated protein kinase

Rac1:

Ras-related C3 botulinum toxin substrate 1

ROS:

Reactive oxygen species

T2DM:

Type 2 diabetes mellitus

ZDF:

Zucker diabetic fatty rat

ZLC:

Zucker lean control

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Acknowledgements

This work was supported by grants to A.K from the Department of Veterans Affairs (Merit Review Program) and National Institutes of Health, and a Research Stimulation Award from Wayne State University. A.K is the recipient of a Senior Research Career Scientist Award from the Department of Veterans Affairs (13S-RCS-006). VS received a Pre-Doctoral Fellowship from Graduate School at Wayne State University. The authors thank Drs. Anil K. Chekuri and Anil Poudel for contributions and support throughout these studies.

Author contributions

Vaibhav Sidarala performed the experimental work, analyzed the data and edited the manuscript prior to publication. Anjan Kowluru designed the studies, wrote the manuscript and edited prior to submission.

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Author notes

  1. Vaibhav Sidarala

    Present address: Division of Metabolism, Endocrinology and Diabetes (MEND), Department of Internal Medicine, Brehm Diabetes Research Center, University of Michigan Medical School, Ann Arbor, MI, 48105-1912, USA

Authors and Affiliations

  1. Beta-Cell Biochemistry Laboratory, John D. Dingell VA Medical Center, B-4237 Research Service, 4646 John R, Detroit, MI, 48201, USA

    Vaibhav Sidarala & Anjaneyulu Kowluru

  2. Department of Pharmaceutical Sciences, Wayne State University, Detroit, MI, 48201, USA

    Vaibhav Sidarala & Anjaneyulu Kowluru

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  1. Vaibhav Sidarala
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Correspondence to Anjaneyulu Kowluru.

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Sidarala, V., Kowluru, A. Exposure to chronic hyperglycemic conditions results in Ras-related C3 botulinum toxin substrate 1 (Rac1)-mediated activation of p53 and ATM kinase in pancreatic β-cells. Apoptosis 22, 597–607 (2017). https://doi.org/10.1007/s10495-017-1354-6

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